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Microparticle-tagged image-based cell counting (ImmunoSpin) for CD4 + T cells.

Sang-Hyun Hwang1,2, John Jeongseok Yang1, Yoon-Hee Oh1

  • 1Department of Laboratory Medicine, Asan Medical Center, University of Ulsan College of Medicine, Seoul, 05505, Republic of Korea.

Mikrochimica Acta
|November 25, 2021
PubMed
Summary

A new ImmunoSpin method accurately counts CD4+ T lymphocytes using microparticles and a light microscope, offering a precise point-of-care alternative for HIV patients. This visual detection method avoids complex systems and shows high correlation with flow cytometry.

Keywords:
CD4 + T cellHuman immunodeficiency virusImage-based cell countingImmunoSpinMicroparticle

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Area of Science:

  • Immunology
  • Biotechnology
  • Medical Diagnostics

Background:

  • Accurate CD4+ T lymphocyte counting is crucial for managing HIV-1 infection.
  • Existing point-of-care (POC) CD4 enumeration technologies can lack accuracy compared to flow cytometry.
  • There is a need for affordable, reliable POC CD4 testing methods.

Purpose of the Study:

  • To develop and validate ImmunoSpin, a novel microparticle-based visual detector for CD4+ T lymphocytes.
  • To assess the accuracy, precision, and reliability of ImmunoSpin as a POC diagnostic tool.
  • To compare ImmunoSpin's performance against standard flow cytometry methods.

Main Methods:

  • Development of microparticles conjugated with anti-CD4 antibodies for visual detection.
  • Enumeration of CD4+ T lymphocytes using a conventional light microscope.
  • Comparison of ImmunoSpin results with clinical-grade flow cytometry for accuracy and precision.
  • Evaluation of ImmunoSpin's ability to differentiate CD4+ T cells from other blood cells.

Main Results:

  • ImmunoSpin demonstrated accurate visual enumeration of CD4+ T lymphocytes under a light microscope.
  • The method exhibited high precision (CV ≤ 10%) and strong correlation with flow cytometry (R² = 0.99).
  • No misclassification occurred at critical CD4+ thresholds (200 and 350 cells/µL), and clear differentiation from other leukocytes was observed.

Conclusions:

  • ImmunoSpin provides an accurate, precise, and affordable POC method for CD4+ T lymphocyte counting.
  • The non-fluorescence, visual detection system is suitable for resource-limited settings and potential automation.
  • ImmunoSpin offers a reliable alternative to flow cytometry for HIV patient management.